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Simultaneous Immunohistochemical Demonstration of Antigen Expression and 5-Bromodeoxyuridine Incorporation in Plastic Embedded Sections by G. Harms; H. van Goor; J. Koudstaal; L. de Ley; M. J. Hardonk is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

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In this study a double immunohistochemical staining procedure is described for the simultaneous demonstration of antigen expressing cells and replicating cells in rat thymus. As markers for cell surface antigen expression a monoclonal antibody against Ia-expressing cells (His 19) and a monoclonal antibody against cells of the monocytemacrophage lineage (ED2) were used. Replicating cells were demonstrated by the incorporation of 5-bromodeoxyuridine (BrdUrd). Tissue pieces were fixed in a periodate-lysine-paraformaldehyde fixative and embedded in glycol methacrylate. To demonstrate h-expressing cells or ED2positive macrophages in plastic embedded sections a digestion with trypsin is necessary. The staining procedure was applied sequentially and was performed with a peroxidase and an alkaline phosphatase labeled reagent yielding respectively a brown and a blue reaction product. Results with this staining procedure on plastic embedded sections of rat thymus, an organ with a high DNA synthesizing capacity, showed incorporation of BrdUrd predominantly in the cortex. ED2-positive macrophages were only found in the cortex. The Ia-positive epithelial reticular cells demonstrated extremely w

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Author

G. Harms; H. van Goor; J. Koudstaal; L. de Ley; M. J. Hardonk

Publisher

Springer; Springer-Verlag; Springer Verlag; Springer Science and Business Media LLC (ISSN 1432-119X)

Published

1987

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)